L‐arginine‐nitric oxide pathway involvement in the nerve‐evoked relaxant responses of the 5‐HT precontracted chick isolated upper oesophagus

Abstract

1. The effects of tetrodotoxin (TTx) and the selective nitric oxide synthase (NOS) inhibitor, NG-nitro-L-arginine methyl ester (L-NAME) on relaxant responses of the 5-HT precontracted chick isolated upper oesophagus to electrical field stimulation (EFS: 25-30V, 5 Hz for 10 s, 1 ms pulse width every 100 s) were investigated; the oesophagus was mounted under 1 g tension in Krebs solution containing 1 microM atropine. Appropriate tissue sections (30 microM thickness) of the chick oesophagus were also processed for NADPH-diaphorase histochemistry. 2. TTx (2 microM) and L-NAME (100-200 microM) inhibited the relaxant responses of the 5-HT precontracted chick oesophagus to EFS in a concentration-dependent manner; L-arginine (0.5-1 mM), but not D-arginine (0.5-1 mM), reversed the inhibition by L-NAME. In the absence of atropine and muscle tone, EFS produced contractile responses of the chick oesophagus that were completely abolished by 1 microM atropine, which also blocked the contractile response to acetylcholine (50 microM). 3. Under light microscopy, NADPH-diaphorase histochemistry confirmed the presence of nitric oxide synthase (NOS)-containing neurones and nerve fibres in the chick oesophagus. 4. The relaxant responses of the 5-HT precontracted chick isolated upper oesophagus to EFS are, therefore, mediated via the stimulation of non-adrenergic non-cholinergic nerves. These are likely to correspond to the histochemically identified NOS-containing neurones involved, presumably, in the synthesis and release of nitric oxide as the relaxant (inhibitory) neurotransmitter in this avian smooth muscle.